Total RNA was isolated by using TRIzol reagent (Invitrogen, USA) according to the manufacturer’s instructions. The isolated total RNA was reverse-transcribed by using PrimeScript RT reagent Kit (TaKaRa, Shiga, Japan). Real-time PCR was performed on an ABI 7900 Real-Time PCR System (Applied Biosystems). Specific primers for Real-time PCR were listed in Additional file 5 : Table S1. The gene expression is quantified by normalizing to 18s.
Western Blot analysis was performed as described before [40 (link)]. Anti-SCD-1 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-FASN (BD, Biosciences PharMingen, San Diego, CA, USA), anti-ACC1 (Cell Signaling Technology, Danvers, MA), anti-β-Actin (Sigma), anti-α-Tubulin (Sigma), and anti-GYS2 (Cell Signaling Technology) antibodies were used as primary antibodies.
Western Blot analysis was performed as described before [40 (link)]. Anti-SCD-1 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-FASN (BD, Biosciences PharMingen, San Diego, CA, USA), anti-ACC1 (Cell Signaling Technology, Danvers, MA), anti-β-Actin (Sigma), anti-α-Tubulin (Sigma), and anti-GYS2 (Cell Signaling Technology) antibodies were used as primary antibodies.
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